Mesenchymal Stem/Stromal Cell-Based Delivery: A Rapidly Evolving Strategy for Cancer Therapy.

Mesenchymal stem/stromal cell (MSC)-based therapy has become an attractive and advanced scientific research area in the context of cancer therapy. This interest is closely linked to the MSC-marked tropism for tumors, suggesting them as a rational and effective vehicle for drug delivery for both hematological and solid malignancies. Nonetheless, the therapeutic application of the MSCs in human tumors is still controversial because of the induction of several signaling pathways largely contributing to tumor progression and metastasis. In spite of some evidence supporting that MSCs may sustain cancer pathogenesis, increasing proofs have indicated the suppressive influences of MSCs on tumor cells. During the last years, a myriad of preclinical and some clinical studies have been carried out or are ongoing to address the safety and efficacy of the MSC-based delivery of therapeutic agents in diverse types of malignancies. A large number of studies have focused on the MSC application as delivery vehicles for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), chemotherapeutic drug such as gemcitabine (GCB), paclitaxel (PTX), and doxorubicin (DOX), prodrugs such as 5-fluorocytosine (5-FC) and ganciclovir (GCV), and immune cell-activating cytokines along with oncolytic virus. In the current review, we evaluate the latest findings rendering the potential of MSCs to be employed as potent gene/drug delivery vehicle for inducing tumor regression with a special focus on the in vivo reports performed during the last two decades.

Introduction

There is a rapidly evolving interest in the marked competencies of the cell-based approach for amelioration of a myriad of pathological conditions ranging from degenerative disorders to cancers (Ramdasi et al., 2015; Wang M. et al., 2018). Mesenchymal stem/stromal cells (MSCs) as non-hematopoietic progenitor cells can be procured from various types of human tissues containing bone marrow (BM), adipose tissue (AT), the dental pulp (DP), muscle, skin, placenta, and also umbilical cord blood (UCB) (Majumdar et al., 1998; Almeida-Porada et al., 2020; Tavakoli et al., 2020). The procurement process of MSCs from adult tissues resolves the ethical issues of application of embryonic or fetal stem cells, while still offering cells with the capacity to differentiate into numerous adult cell lineages, surrounding adipocytes, osteocytes, and chondrocytes in vitro (Reger et al., 2008; Lai et al., 2019). Regardless of differentiation into adipocytes, osteocytes, and chondrocytes in vitro, demonstrating plastic adherence possessions concomitant with the expression of CD73, D90, and CD105 and lack of expression of CD14, CD34, CD45, and human leucocyte antigen-DR (HLA-DR) are other criteria offered by the International Society for Cellular Therapy (ISCT) enabling characterization of MSCs in varied cell population (Dominici et al., 2006; Bühring et al., 2007; Krampera et al., 2013; Galipeau and Krampera, 2015). Meanwhile, a unique characteristic of MSCs is their lower immunogenicity due to the lack of expression of costimulatory molecules (Najar et al., 2012; Wu et al., 2014), which eliminates the need for using immunosuppressive agents postallogeneic transplantation. This characteristic makes them supremely appropriate for cell-based cancer immunotherapy as either gene or drug delivery vehicles (Le Blanc and Ringden, 2006; Kean et al., 2013; Nasr et al., 2015). Furthermore, their comparative ease of harvest from a diverse sources, preservation of the functionality during cultivation in culture, displaying immunoregulatory aptitudes, and also remarkable efficacy once modified with viral-based vectors along with their evident potential to specifically home to impaired tissue upon systemic injection highlight advantages of MSC in the context of cancer cell-based therapy (Sung et al., 2008; Kean et al., 2013; Almeida-Porada et al., 2020).

Currently, substantial efforts have been made to employ the natural capability of MSC for recruitment to inflammatory areas such as those existing in tumors (Hmadcha et al., 2020). Indeed, MSC-based therapies are encouraging strategy for the advancement of gene transfer systems and drug-loading approaches due to their intrinsic properties, containing homing ability concomitant with tumor tropism. By merging the inherent cell competencies and antitumor capacities of cytokines, utilizing MSC modified to express target cytokines or other ingredients results in more efficient cytokine/drug delivery to tumor site, providing a paradigm shift in the context of tumor immune therapy (Wu et al., 2019). For example, MSCs genetically modified to express interleukin-12 (MSC-IL-12) demonstrated more prominent tumor-specific T-cell responses and anticancer impacts, and also more sustained expressions of IL-12 and interferon (IFN)-γ in both sera and tumor sites than IL-12-expressing viral vector therapy in murine with metastatic tumors (Seo S.H. et al., 2011). In addition, MSCs loaded with oncolytic virus (OV) could dramatically kill tumor cells in vitro and in vivo and provide antitumor effects with improved safety profiles (Yoon et al., 2019). Meanwhile, majority of preclinical reports signify safety and efficacy of MSC employment as carriers of OV without any tumorigenicity (Hadryś et al., 2020). Furthermore, MSC-based delivery of the tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a potent antitumor molecule, could restore the sensitivity of TRAIL-resistant tumor cells to TRAIL-induced apoptosis (Rossignoli et al., 2019). Accordingly, it has been evidenced that MSCs can be exploited to serve a range of therapeutic ingredients, comprising apoptosis-inducing ingredient (e.g., TRAIL), chemotherapeutic agents (e.g., gemcitabine, paclitaxel, and doxorubicin), tumor- or tissue-specific prodrugs (e.g., 5-fluorocytosine and ganciclovir), immune cell-inducing cytokines (e.g., interleukin and interferon), and oncolytic virus (e.g., herpes simplex virus, measles virus, and adenovirus) (Figure 1; Loebinger et al., 2009; Kerrigan et al., 2017; Krueger et al., 2018; Lisini et al., 2020). Herein, we will deliver a brief overview of the latest research respecting MSC application as gene/drug delivery vehicles for supporting tumor cell elimination and averting tumor growth, focusing on the in vivo reports from last two decades.

FIGURE 1

MSC-based delivery for cancer treatment. Human MSCs can be utilized to stimulate tumor regression following isolation and modification, acting as a vehicle for the delivery of drugs or molecules into tumors. MSCs, mesenchymal stem/stromal cells; TRAIL/Apo2L, tumor necrosis factor (TNF)-related apoptosis-inducing ligand.

MSC-Based Delivery of Cytokines

Proinflammatory cytokines play a role in every stage of the cancer immunity cycle and can be used for tumor immunotherapy. They can mend antigen priming, promote the frequency of effector immune cells in the tumor microenvironment (TME), and also boost their cytolytic functions. In this section, we have reviewed the recent reports respecting MSC-based cytokine delivery for cancer therapy (Table 1).

TABLE 1

MSC-based delivery of cytokines for cancer therapy.

Cancer	Cytokine	Main results
Melanoma	IL-2	Induction of CD8-associated tumor-specific immunity and inhibition of tumor development in B16 cell-bearing mice by IL-2-producing MSCs (Stagg et al., 2004)
Glioma	IL-2	Hindrance of tumor growth and promotion of the overall survival of 9L glioma-bearing rats by intratumoral administration of IL-2-producing MSCs (Nakamura et al., 2004)
Melanoma Cervical cancer	IL-12	Antitumor effects in melanoma and cervical bearing mice by administration of IL-12-producing MSCs embedded in Matrigel (Seo S.H. et al., 2011)
Renal cell carcinoma (RCC)	IL-12	Attenuation of the growth of 786-0 RCC and also improvement of the RCC mouse models survival by activation of NK cells and IFN-γ secretion by systemic administration of IL-12-producing MSCs (Gao et al., 2010)
Breast cancer	IL-12	Induction of the elimination of 4T1 cells concomitant with induction of antiangiogenesis effects by IL-12-producing MSCs (Eliopoulos et al., 2008)
Melanoma Breast cancer Hepatoma	IL-12	Induction of the tumor cell elimination in B16 melanoma, 4T1 breast tumor, and HCA hepatoma cancer models by IL-12-producing MSCs (Chen et al., 2008)
Glioma	IL-12	Inhibition of tumor growth and extended survival of glioma-bearing mice by injection of IL-12-producing UCB-MSC (Ryu et al., 2011)
Ovarian cancer	IL-21	Promotion of IFN-γ-secretion and NK cells cytotoxicity resulted in inhibition of tumor growth in ovarian cancer-bearing mice by injection of IL-21-producing UCB-MSC (Hu et al., 2011)
Lymphoma	IL-21	Delayed cancer progress and promoted survival of lymphoma-bearing mice mediated by stimulation of effector T and NK cells activities upon injection of IL-21-producing MSC (Kim et al., 2015)
Lung carcinoma	IFN-γ	Inhibition of growth and progression of lung carcinoma by activation of TRAIL pathways in nude mice following injection of the MSC producing IFN-γ (Yang et al., 2014)
Melanoma	IFN-α	Delayed development of B16F10 melanoma cells led to the prolonged survival of tumor-bearing C57BL/6 mice (Ren et al., 2008a)
Breast cancer	IFN-β	Suppression of the breast tumor growth and reduction of pulmonary and hepatic metastases in tumor-bearing mice along with down-regulation of Stat3, Src, and Akt cMyc and MMP2 expression by systemic injection of MSC producing IFN-β (Ling et al., 2010)
Prostate cancer lung metastasis	IFN-β	Attenuation in tumor volume in lungs after injection of MSC producing IFN-β and promoted tumor cell apoptosis and reduced tumor cell proliferation (Ren et al., 2008b)
Pancreatic cancer	IFN-β	Suppression of tumor growth and downregulation of NF-κB, VEGF, and IL-6 in orthotopic PANC-1 pancreatic carcinoma SCID mice models following systemic injection of MSC producing IFN-β (Kidd et al., 2010)
Bronchioloalveolar carcinoma	IFN-β	Attenuation of the development of orthotopic H358 bronchioloalveolar carcinoma in SCID mice through stimulating apoptosis by systemic injection of MSC producing IFN-β (Matsuzuka et al., 2010)
Lung carcinoma	IFN-β	Delayed tumor growth in non-small cell lung cancer (NSCLC) mouse model by systemic injection of UC-MSC producing IFN-β (Chen et al., 2019)
Squamous cell carcinoma (SCC)	IFN-β	Delayed tumor growth in SCC xenograft by administration of GMSCs producing IFN-β in BALB/c nude mouse model (Du et al., 2019)
Melanoma	IFN-β	Inhibition of the progress of melanoma and extended survival of transplanted canine models by the combination treatment of AT-MSC producing IFN-β with cisplatin (Ahn et al., 2013)

Interleukins

In 2004, Nakamura et al. found that MSCs in vivo could migrate into tumor area in 9-L glioma-bearing murine models through the corpus callosum and finally infiltrate into the tumor bed (Nakamura et al., 2004). Intratumoral administration of MSCs significantly hindered 9-L tumor growth and markedly augmented the survival of 9-L glioma-bearing murine (Nakamura et al., 2004). Importantly, administration of engineered MSCs by adenoviral vector expressing human IL-2 improved the antitumor influences and further extended the overall survival rates of tumor-bearing murine, delivering the preliminary proof of the concept indicating MSC competencies to be employed as cytokine delivery vehicles (Nakamura et al., 2004). Similarly, BM-MSCs modified to overexpress IL-2 were found to delay tumor growth in B16 melanoma-bearing murine models, while parental BM-MSCs did not have any inhibitory effects on tumor development in the transplanted models (Stagg et al., 2004). Moreover, administration of the matrix-embedded IL-2-producing MSCs into B16 melanoma-bearing models resulted in inhibition of tumor growth by CD8 and Natural Killer (NK) cells, but not CD4 cell activation (Stagg et al., 2004). In addition, human AT-MSC-producing IL-2 were suggested to release a higher rate of proinflammatory cytokines compared with parental stem cells, and also supported peripheral blood mononuclear cell (PBMCs) activation and conversely averted the viability of SH-SY5Y neuroblastoma cells in co-culture condition in vitro (Chulpanova et al., 2020). Nonetheless, the observation signified that IL-2-mediated therapeutic impacts of AT-MSCs could be impaired by the boosted expression of pro-oncogenes, accompanied by the innate capability of AT-MSCs to develop some tumors (Chulpanova et al., 2020).

In addition to IL-2, injection of MSC-producing IL-12 leads to promising results in the animal models. Meanwhile, reports signify that intravenous, intratumoral, and subcutaneous administration of MSC-producing IL-12 could display more powerful tumor-specific T-cell responses and anticancer effects than the injection of parental MSCs in both B16F10 melanoma and TC-1 cervical tumor-bearing mice, whereas the antitumor effect elicited by cell intratumoral administration was more prominent than subcutaneous and also intravenous injection in both models (Seo S.H. et al., 2011). Also, MSC-producing IL-12 embedded in Matrigel (MSC-IL-12-Matrigel) exhibited substantial antitumor effects in immunodeficient mice comprising severe combined immunodeficient mice (SCID) and beige/nude/X-linked immunodeficient (BNX) mice without T and B lymphocytes (T and B cells) and NK cells but not in interferon-gamma (IFN-γ) knockout mice (Seo S.H. et al., 2011). In addition, administration of MSCs engineered to overexpress IL-12 in B16F10 melanoma-bearing mice models intensely condensed the melanoma lung metastases by induction of NK cell proliferation and activation (Elzaouk et al., 2006). Furthermore, IL-12-producing MSCs caused pronounced retardation of tumor development and resulted in improved survival following subcutaneous injection into tumor-bearing mice (Elzaouk et al., 2006). Besides, systemic injection of BM-MSCs transduced with a recombinant adenoviral vector expressing the murine IL-12 in a 786-0 renal cell carcinoma (RCC)-bearing mice supported the delayed tumor growth and significantly sustained mouse survival following homing to tumors and production of local IL-12 (Gao et al., 2010). The analysis presented that the observed antitumor effects were in association with the presence of NK cells and IFN-γ in animal models (Gao et al., 2010). In addition to the evidenced therapeutic efficacy of IL-12-producing BM-MSCs in Ewing sarcoma tumors (Duan et al., 2009), another study revealed their antitumor effects in murine metastatic hepatoma established by HCA-I and Hepa 1–6 cells. Upregulation of monocyte chemoattractant protein-1 (MCP-1/CCL2) and improved activation of cytotoxic T lymphocytes (CTL) and NK cells in tumor tissue was described as underlying mechanisms largely contributed to the modified MSC-exerted antitumor effects (Jeong et al., 2015). Moreover, treatment inhibited pulmonary metastasis and promoted survival rate in tumor-bearing models by induction of apoptosis of tumor cells through stimulation of the effector immune cell proliferation (Jeong et al., 2015).

Besides, current investigations have indicated that IL-21 delivery using MSCs could lead to the antitumor effects in a variety of cancers by affecting immune cell activation and proliferation. For instance, systemic injection of IL-21-producing UCB-MSCs in A2780 ovarian cancer-bearing mice models boosted IFN-γ-generating splenocyte numbers and NK cell cytotoxicity in transplanted models (Hu et al., 2011); on the other hand, intervention suppressed tumor development and enhanced the survival rate of tumor-bearing mice (Hu et al., 2011). Likewise, injection of IL-21-secreting human UC-MSCs into SKOV3 ovarian tumor-bearing nude mice potently attenuated tumor burden in transplanted models, as shown by reduced tumor sizes (Zhang et al., 2014). Importantly, IL-21 delivery modified the levels of IFN-γ and tumor necrosis factor-alpha (TNF-α) in the mouse serum, augmented natural Killer group 2D (NKG2D) and MHC class I polypeptide-related sequence A (MICA) molecule expressions in the tumor tissues, and finally downregulated β-catenin and cyclin-D1 in the tumor, which in turn, led to delayed tumor growth posttransplantation (Zhang et al., 2014). Similarly, IL-21-overexpressing MSCs were found to sustain antitumor responses by local delivery of IL-21 in B-cell lymphoma BALB/c mice models, thus preventing the establishment of tumor nodules (Kim et al., 2015). Although the IL-21-MSC-treated group presented reduced tumor development and enhanced survival by induction of effector T and NK cells; administration of the MSCs and recombinant adenovirus-expressing IL-21 (rAD/IL-21) had no significant antitumor effects in transplanted models (Kim et al., 2015).

Interferons

Based on the literature, adult stem cells show notable potential for delivery of IFNs to tumor tissues to induce the antitumor functions of immune cells (Ahn et al., 2013). For instance, Ren et al. (2008a) found that interferon-alpha (IFN-α)-overexpressing BM-MSCs could exert suppressive effects on lung metastasis in B16F10-bearing C57BL/6 mice model of metastatic melanoma. Accordingly, systemic infusion of IFN-α-producing MSC abridged the tumor growth and sustained survival in experimental models. Immunohistochemistry analysis verified an enhancement in apoptosis and a reduction in proliferation and blood vasculature, representing the capability of adult IFN-α-overexpressing MSC in diminishing the progress of lung metastasis in melanoma (Ren et al., 2008a). Another study showed that even a small population of MSCs constitutively generating IFN-α could strongly delay B16 tumor development in xenograft models mediated by activation of NK and CD8-positive T cells (Xu et al., 2014).

The incorporation of other interferons with MSCs has also been explored by researchers. Yang et al. suggested that MSC constitutively producing IFN-γ could eliminate tumor cells by continued activation of the apoptosis-inducing TRAIL pathway (Yang et al., 2014). Correspondingly, IFN-γ-modified MSCs could strongly release functional IFN-γ, leading to the prolonged expression of TRAIL and subsequent activation of caspase cascade in tumor cells. For example, IFN-γ-secreting MSCs selectively triggered apoptosis in lung tumor cells via upregulation of caspase-3 activation following improvement of TRAIL expression in tumor cells in vitro. Moreover, in xenograft mice models, MSC constitutively generating IFN-γ hindered the progress of lung carcinoma (Yang et al., 2014). In another research, MSCs generating IFN-γ polarized murine macrophages to the M1 phenotype in vitro, and also delayed tumor growth supporting improved overall survival in neuroblastoma tumor cell-bearing xenografts (Relation et al., 2018). Besides, co-culture of IFN-γ-overexpressing BM-MSCs with human chronic myelogenous leukemia (CML) K562 cells led to the potent suppression of the leukemic cell proliferation along with their apoptosis in the experimental groups as well as induction of cell cycle arrest at G1 phase (Li et al., 2006).

In addition to the IFN-α and IFN-γ gene delivery by MSCs, MSC-producing interferon beta (IFN-β) have represented more effective antitumor effects. Studies in breast tumor xenografts revealed that intravenously injected MSC could migrate to the breast tumor area and secrete high rates of IFN-β into tumor stroma (Ling et al., 2010). Molecular analysis demonstrated that intratumorally generated IFN-β could downregulate activation of signal transducer activator transcription factor 3 (Stat3), Src, Akt, cMyc, and matrix metalloproteinase-2 (MMP-2) expression in tumor cells which led to the suppression of primary tumor growth and attenuation of pulmonary and hepatic metastases (Ling et al., 2010). Moreover, assessment of the antitumor effects of combination therapy with the canine AT-MSCs producing IFN-β and cisplatin in B16F10 melanoma-bearing mice evidenced that intratumoral administration of cisplatin along with subcutaneous injection of modified AT-MSCs was superior in terms of abrogation of the melanoma tumor growth and survival, over the administration of modified AT-MSCs or cisplatin alone (Seo K.W. et al., 2011). Also, promising results achieved from the treatment of human U87 glioma-bearing mice models with IFN-β-secreting MSCs signified that MSCs could be recruited into the brain tissue after systemic or local delivery, thereby enabling human glioma treatment (Nakamizo et al., 2005). Besides, administration of the IFN-β-overexpressing MSCs caused a significant reduction in the tumor volume in prostate cancer lung metastasis in xenograft model (Ren et al., 2008b) and also squamous cell carcinoma xenograft model (Du et al., 2019).

MSC-Based Oncolytic Virus Delivery

Virotherapy as an innovative strategy for cancer treatment has some advantages such as the potential absence of cross-resistance with conventional therapies and also the capacity to induce tumor eradication by several mechanisms. Regardless of their unique mechanisms of functions distinguishing them from other treatment options, the self-perpetuating nature of oncolytic virus (OVs) serves as a supreme strategy for therapeutic transgenic insertion. The use of MSCs as a non-systemic carrier of OVs has been investigated with remarkable achievement for the treatment of varied types of pathological conditions (Figure 2). In this section, the potent antitumor effects of the OVs following delivery by MSCs into tumor tissues is reviewed with emphasis on AV, HSV, and MV (Table 2).

FIGURE 2

MSC-based delivery of oncolytic virus (OV). Though unshielded OV commonly stimulates antiviral response following systemic administration, which results in virus clearance without any oncolytic action, MSC-based delivery of OV enables effective delivery of virus to tumor stroma and subsequent oncolytic action. MSCs, mesenchymal stem/stromal cells; IV, intravenous.

TABLE 2

MSC-based delivery of oncolytic viruses for cancer therapy.

Cancer	OV	Main results
Hepatocellular carcinoma (HCC)	AV	Potent tumor growth inhibition in HCC murine models by MSC-mediated delivery of oAV (Yoon et al., 2019)
Glioblastoma (GBM)	HSV AV	The killing of GBMs in vitro and in vivo mediated by MSC-mediated delivery of oHSV and oAV resulted in promoted GBM cell eradication and prolonged median survival in the mice model (Duebgen et al., 2014; Leoni et al., 2015)
Brain metastatic melanomas	HSV	Prolonged survival of brain tumor-bearing mice by MSC-mediated delivery of oHSV (Du et al., 2017)
Lung and brain metastatic ovarian cancer	HSV	Inhibition of the growth of ovarian cancer lung metastases in nude mice by MSC-mediated delivery of oHSV (Leoni et al., 2015)
Ovarian cancer	MV	Promotion of the survival of the measles-immune ovarian tumor mice models after MSC-mediated delivery of oMV (Mader et al., 2009)
Hepatocellular carcinoma (HCC)	MV	Remarkable obstruction of tumor growth in both measles antibody-naïve and passively immunized SCID HCC mice models by BM-MSC-mediated delivery of oMV (Ong et al., 2013)
Acute lymphocytic leukemia (ALL)	MV	Inhibition of cancer development in a murine model of disseminated ALL following MSC-mediated delivery of oMV (Castleton et al., 2014)
Lung cancer Breast cancer	AV	Delayed progress of the orthotopic breast and lung tumors caused increased survival of murine models by MSC-mediated delivery of oAV (Hakkarainen et al., 2007)
Pancreatic cancer	AV	Remarkable antitumor effect in pancreatic tumor model by MSC-mediated delivery of oAV (Hammer et al., 2015)
Hepatocellular carcinoma (HCC)	AV	Significant tumor suppression in both orthotopic and subcutaneous hepatic tumor mice model by MSC-mediated delivery of oAV (Yuan et al., 2016)
Lung carcinoma	AV	Improvement of the tumor infiltration of CD8+ and CD4+ T cells in lung carcinoma murine models by MSC-mediated delivery of oAV (Rincón et al., 2017)
Neuroblastoma	AV	Augmented infiltration of immune cells resulted in a reduction in tumor mass in vivo by MSC-mediated delivery of oAV (García-Castro et al., 2010)
Metastatic breast cancer	AV	Prolonged mouse survival with MDA-MB-231-derived pulmonary metastatic tumor by MSC-mediated delivery of oAV (Stoff-Khalili et al., 2007)
Colorectal cancer (CRC)	AV	Potent inhibitory effects against CRC in mice model by MenSC-mediated delivery of oAV (Guo et al., 2019)
Prostate cancer	AV	Inhibition of tumor growth in subcutaneous human prostate cancer mice models by MSC-mediated delivery of oAV (Park et al., 2010)

Oncolytic Adenovirus

Investigation of the immune response to MSCs loaded with oncolytic adenovirus (MSC-oAd) using the semipermissive cotton rat (CR) model verified that CR-MSCs could sustain the replication of oncolytic adenovirus (oAd) in vitro (Ahmed et al., 2010). Furthermore, CR-MSCs could inhibit IFN-γ production by activated T cells and more intensely promote the distribution and persistence of oAd in comparison with virus injection alone in vivo. This indicates that MSC employment as a delivery tool for oAd could provide several merits, such as supported delivery, promoted distribution, and ameliorated persistence of viruses by inhibition of the antiviral immune reactions (Ahmed et al., 2010). Furthermore, intravenously injected MSC-oAd could home to hepatocellular carcinoma (HCC) tumors in xenograft models and support promoted rates of the virion gathering in the tumors, causing marked tumor growth inhibition in experimental models (Yoon et al., 2019). In addition, MSC-oAd potently killed HCC cells in vitro under both normoxic and hypoxic conditions, indicating that MSC-associated systemic delivery of oAd is a promising strategy for achieving synergistic antitumor efficacy with improved safety profiles (Yoon et al., 2019). Similarly, MSC-oAd showed antitumor effects in orthotopic murine models of lung and breast tumor following systemic injection which resulted in improved survival of xenograft models, while injection of unshielded oAd without the use of MSCs only led to the transduction of the liver (Hakkarainen et al., 2007).

Systemic administration of human MSC transduced with conditionally replicating adenoviruses (CRAds) in a SCID mouse xenograft metastases model of breast tumors resulted in effective recruitment of injected cells into tumor area in experimental models and consequently improved mouse survival in comparison with mice treated with CRAds alone apparently by viral amplification in the MSCs (Stoff-Khalili et al., 2007). On the other hand, Guo and his colleagues found that menstrual blood (Men)-derived MSCs loaded with oAd could display robust suppressive impacts on colorectal cancer (CRC) growth in mice models possibly by viral amplification, as evidenced by higher rates of the viruses accumulated in tumor site (Guo et al., 2019). In addition to the induction of suppressive effects on the proliferation of prostate cancer cells in vitro, MSC-oAd demonstrated tumor tropism in prostate cancer mice models concomitant with suppression of tumor growth in experimental models. This result arose from continued viral replication which enabled cell lysis (Muhammad et al., 2019).

In 2010, evaluation of the safety and the efficacy of injection of autologous MSCs transduced with ICOVIR-5, a new oAd, in four children with metastatic neuroblastoma demonstrated that the injection was well tolerated; on the other hand, a complete clinical response was verified in one participant, describing MSCs as eligible vehicles for oncolytic virus delivery into metastatic tumors with very low systemic toxicity concomitant with reliable antitumor effects (García-Castro et al., 2010).

Oncolytic Herpes Simplex Virus

It has been found that MSCs loaded with oncolytic herpes simplex virus (oHSV) (MSC-oHSV) could potently generate oHSV progeny leading to the lysis of glioblastoma (GBM) cells in vitro and in vivo presumably by a dynamic procedure of oHSV infection and tumor eradication (Duebgen et al., 2014). Moreover, biocompatible synthetic extracellular matrix (sECM)-encapsulated MSC-oHSV could trigger improved anti-GBM efficacy in comparison with the direct use of purified oHSV in the xenograft model, causing extended survival in animal models (Duebgen et al., 2014). Furthermore, MSC loaded with oHSV-TRAIL could robustly stimulate apoptosis-related killing and prolonged survival in mice bearing oHSV- and TRAIL-resistant GBM (Duebgen et al., 2014). In addition, MSCs infected with a human epidermal growth factor receptor 2 (HER2)-retargeted oHSV resulted in the spreading of the oHSV progeny from MSCs to tumor cells in both metastatic breast and ovarian cancer murine models upon systemic injection (Leoni et al., 2015). Observation supported the existence of the remarkable concentration of MSCs and viral genomes in the lungs of experimental models and also showed the reduced tumor growth in nude mice in the absence of any significant metastasis in the majority of treated mice (Leoni et al., 2015).

Study of the antitumor effects of the MSCs loaded with HF10, a HSV-1 mutant, in combination with tyrosine kinase inhibitor erlotinib in vitro and pancreatic tumor-bearing murine models demonstrated that combination therapy led to the highest cytotoxicity toward human pancreatic cell line BxPC-3, while combined use of MSC-HF10 and erlotinib had no significant cytotoxicity against human pancreatic cell line PANC-1 cells (Yamamura et al., 2014). Significantly, the combined use of MSC-HF10 and erlotinib supported persistent virus propagation in tumor areas accompanying with suppression of tumor development more prominent than using each of them alone in the subcutaneous tumor model (Yamamura et al., 2014).

Oncolytic Measles Virus

It has already been suggested that the use of human MSCs loaded with oncolytic measles virus (oMV) (MSC-oMV) can lead to the desired therapeutic outcomes in a variety of human tumors (e.g., ovarian tumors) (Mader et al., 2009). In this regard, studies in SKOV3ip.1 ovarian tumor xenografts demonstrated that intraperitoneally injected MSC could home to peritoneal tumors, migrate into the tumor parenchyma, and finally convey virus infection to tumors in both measles-naïve and passively immunized mice. Remarkably, administration of the MSC-oMV but not a naked virus or uninfected MSC resulted in improved survival of orthotopic ovarian cancer models (Mader et al., 2009). In addition, it has been found that patient-derived MSC can be preinfected with oMV, stored in liquid nitrogen, and finally thawed before administration without losing their functionality (Mader et al., 2013). Moreover, systemic injection of the BM-MSC-oMV into human HCC SCID mice models supported potent obstruction of tumor growth in both measles antibody-naïve and passively immunized SCID mice. However, injection of naked MV viruses elicited antitumor responses only in measles antibody-naïve SCID mice, which indicated the feasibility and efficacy of MSC-oMV application in treating human liver tumors (Ong et al., 2013). Regardless of the promising outcomes observed in solid tumors, MV delivery using MSCs hold promise for the treatment of hematological malignancies. Accordingly, systemic administration of the BM-MSC-oMV in xenograft model of ALL (Castleton et al., 2014) and also MM (Marchica et al., 2016) stimulated antitumor effects including inhibition of cancer development by induction of malignant cell apoptosis leading to the improved survival in transplanted models.

MSC-Based Chemotherapeutic Agent Delivery

The partial resistance of MSC to cytostatic and cytotoxic chemotherapeutic agents along with their suitable tropism to tumors offers a novel platform for direct targeted delivery of therapeutic agents, such as paclitaxel (PTX), gemcitabine (GCB), and doxorubicin (DOX) to tumor tissue (Table 3). Regardless of the direct application of MSC, extracellular vesicles (EVs) derived from MSCs serve as a substitute method for delivering chemotherapeutic agents.

TABLE 3

MSC-based delivery of chemotherapy agents for cancer therapy.

Cancer	Agent	Main results
Glioblastoma	PTX	Inducing the elimination of human pancreatic carcinoma (CFPAC-1) and glioblastoma (U87-MG) cell line in combination therapy with MSC-TRAIL in vitro (Coccè et al., 2020)
Pancreatic cancer	PTX	Marked antitumor effects on human pancreatic cell line CFPAC-1by MSC-PTX secretome (Coccè et al., 2019)
Breast cancer	PTX	Suppressing the viability of breast cancer MDA-MB-231 cells in vitro and in xenografts (Kalimuthu et al., 2018a)
Pancreatic cancer	PTX	Sustained and continued releases of PTX resulted in improved cell’s eradication (Brini et al., 2016)
Breast cancer	PTX	Stimulation of potent cytotoxicity against MCF-7 cell line in vitro (Salehi et al., 2018)
Prostate cancer	GCB	Inducing anti-proliferative effects on human pCa cell line in vitro (Bonomi et al., 2015)
Squamous cell carcinoma	PTX DXR GCB	Inducing a dramatic suppression of SCC154 cell line proliferation in vitro (Coccè et al., 2017)
Mesothelioma	PTX	Inhibiting the mesothelioma NCI-H28 cell proliferation (Petrella et al., 2017)
Glioma	DXR	Inducing anti-tumor effects in U251 glioma tumor cells bearing murine with broader distribution and prolonged retention lifetime compared with free DOX (Li et al., 2011)
Thyroid cancer Breast cancer	DXR	The pronounced tumor cell elimination in vitro and successful migration to tumor sites in xenografts by MSC-PTX (Kalimuthu et al., 2018b)
Multiple myeloma	PTX	Suppressing the myeloma RPMI 8226 cell proliferation in 3D cell culture system (Bonomi et al., 2017)
Lung carcionoma	PTX	Remarkable anti-tumor effects in orthotopic Lewis lung carcinoma (LL/2-luc) tumors established in C57BL/6 mice (Layek et al., 2018)
Ovarian cancer	PTX	Inhibition of ovarian cancer cells viability in 2D models and in 3D heterospheroids (Borghese et al., 2020)
Leukemia Osteosarcoma Prostate cancer Neuroblastoma	PTX	Potent cytotoxicity against MOLT4 (leukemia), SK-ES-I (anaplastic osteosarcoma), DUl45 (prostatic carcinoma), and GILI-N and SH-SY5Y (neuroblastoma) cell line in vitro (Bonomi et al., 2013)
Osteosarcoma	DXR	Marked antitumor effect against osteosarcoma MG63 cell line but low cytotoxicity in myocardial H9C2 cell line by MSC-exosome-PTX (Wei H. et al., 2019)
Pulmonary metastatic melanomas	DXR	Pronounced antitumor effects in pulmonary B16F10 melanoma metastases in vivo (Zhao et al., 2017)

Paclitaxel

For cell therapy application, a substantial number of MSCs loaded with the antitumor drug is required. Accordingly, some closed bioreactor systems have currently been developed for establishing a large amount of good manufacturing practice (GMP)-compliant MSCs loaded with the chemotherapeutic drug paclitaxel (PTX) (MSC-PTX). In addition to allowing large-scale cultivation of the MSC-PTX, these modified cells show common characteristics of MSCs concerning viability, adhesion capacity, and also phenotype (Lisini et al., 2020). Moreover, MSC-PTX could efficiently internalize PTX and eventually lyse cancer cells and suppress their proliferation in vitro (Lisini et al., 2020). In vivo reports revealed that MSC-based delivery of the TRAIL and PTX in the TRAIL-resistant model of pancreatic cancer could restore sensitivity of tumor cell to MSC-delivered TRAIL by suppression of the expression of survival-related genes (Rossignoli et al., 2019). Furthermore, co-culture of the PTX-primed BM-MSCs with human malignant pleural mesothelioma (MPM), a rare fatal asbestos-related malignancy, and NCI-H28 cells in vitro showed that PTX-loaded BM-MSCs could intensely suppress NCI-H28 cell proliferation; however, in vivo studies are requisite for confirming these preliminary in vitro results (Petrella et al., 2017). Besides, co-culture of the AT-MSCs loaded with PTX with CG5 breast cancer cell line led to the mitigation of the cell proliferation and significant improvement in these cell apoptosis (Scioli et al., 2018). Furthermore, in CG5-bearing murine models, both AT-MSC-PTX and conditioned medium derived from these cells elicited cytotoxic effects against tumor cells (Scioli et al., 2018). Moreover, AT-MSCs primed with paclitaxel could suppress ovarian cancer spheroid development and also defeat paclitaxel resistance (Scioli et al., 2018). Correspondingly, co-culture of ovarian cancer cells with AT-MSC-PTX mitigated cell viability in 2D models and 3D heterospheroids and also lessened PTX-resistance in ovarian cancer Kuramochi cells. In addition, conditioned medium derived from AT-MSC-PTX robustly reduced cancer cell proliferation more prominently than free PTX and attenuated PTX resistance in Kuramochi cells (Scioli et al., 2018; Borghese et al., 2020). Similarly, conditioned medium derived from dental pulp stem cell (DPSC)-PTX showed remarkable cytotoxicity toward human breast cancer MCF-7 cells in vitro following PTX secretion by DPSCs (Salehi et al., 2018). The analysis showed that DPSCs were more resistant to PTX in comparison with BM-MSCs, thus verifying potent capacities of dental stem cell for targeted treatment of cancer (Salehi et al., 2018).

Investigation of the possible antitumor effects of the MSC-PTX encapsulated poly (d,l-lactide-co-glycolide) (PLGA) nanoparticles (NPs) in glioma xenograft rat models clarified superiority of the PTX-PLGA NP-primed MSCs over PTX-primed MSCs in terms of secretion of PTX in the form of free PTX as well as PTX-NPs, supporting glioma cells apoptosis in vitro (Wang X. et al., 2018). Moreover, the employment of the PTX-PLGA NP-loaded MSCs resulted in prolonged survival upon contralateral implantation compared with those injected with PTX-primed MSCs or PTX-PLGA NPs alone which showed that the chemotherapeutic drug-loaded NP incorporation into adult stem cells might be a capable approach for tumor-targeted therapy (Wang X. et al., 2018).

Coccè et al. (2017) evaluated the ability of human MSCs isolated from gingival papilla (GinPa-MSCs) in uptaking and secreting three imperative chemotherapeutic agents, including PTX, DOX, and GCB in co-culture condition with human squamous cell carcinoma SCC154 cells. Based on observations, GinPa-MSCs proficiently secreted agents in active shape and inadequate amount to sustain a notable abrogation of SCC154 cell growth in vitro. Particularly, GCB-primed GinPa-MSCs offer a reliable cell-mediated drug delivery policy for upcoming possible application in oral oncology (Coccè et al., 2017). In addition, PTX-primed hMSCs and mouse bone marrow stromal SR4987 cells (hMSC-PTX and SR4987-PTX) could exert dramatic antitumor effects in MOLT-4 cells bearing mice models as recognized by inhibition of tumor growth as well as angiogenesis, leading to the augmented survival of implanted models (Pessina et al., 2013). Furthermore, both human MSCs and human MSC-PTX secreted molecules which diminished leukemic cell adhesion to microvascular endothelium (MECs) and negatively regulated intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) expression on MECs, revealing the potent competence of this modality for leukemia therapy in humans (Pessina et al., 2013).

Doxorubicin

Currently, studies have shown that doxorubicin (DOX)-primed human BM-MSCs (BM-MSC-DOX) could induce antitumor effects against breast cancer cells (MDA-MB-231) and anaplastic thyroid cancer cells (CAL62) (Kalimuthu et al., 2018b). In vivo, infused BM-MSC-DOX effectively homed to the tumor area in mice thyroid or breast cancer xenografts and then delayed tumor growth. Accordingly, in addition to the evidenced migration of BM-MSC-DOX to tumor stroma in tumor-bearing mice using bioluminescence imaging (BLI), BM-MSC-DOX elicited remarkable cytotoxicity against tumor cells (Kalimuthu et al., 2018b).

In a different design, Liu Y. et al. (2020) generated DOX-loaded superparamagnetic iron oxide (SPIO) nanoparticles (NPs) coated with MSC membranes and evaluated their influence on colon cancer in vitro and in vivo. Compared with DOX-SPIO, MSC membrane-camouflaged nanodrug (DOX-SPIO@MSCs) showed greater cellular uptake efficiency, augmented antitumor influences, and mitigated the immune system reactions in vitro. In addition, DOX-SPIO@MSCs exerted more powerful antitumor effects, while dwindled systemic side effects in MC38 tumor-bearing C57BL/6 mice (Liu Y. et al., 2020). Similarly, silica nanorattle, metallic core-shell particles, -DOX-anchored MSCs were found to track down the U251 glioma cancerous cells more powerfully and also could distribute DOX with wider dissemination and extended retention lifetime in tumor area in comparison with free DOX and silica nanorattle-encapsulated DOX (Li et al., 2011). Correspondingly, silica nanorattle-DOX-anchored MSCs sustained DOX intratumoral disseminations which led to significantly heightened tumor-cell apoptosis (Li et al., 2011). Besides, human umbilical cord (UC)-MSCs carrying transferrin-inspired-nanoparticles containing DOX (hUC-MSC-Tf-inspired-NPs) displayed suitable potential of the secretion of Tf-inspired NPs, and also showed robust tumor tropism (Cao et al., 2018). Importantly, investigations in MCF-7 breast tumor xenograft models indicated that hUC-MSC-Tf-inspired NPs could potently abrogate tumor growth, bringing novel anticancer drug delivery approach (Cao et al., 2018). Remarkably, other reports have shown that injection of DOX-loaded exosomes (Exo) derived from MSCs into an ectopic model of C26 (mouse colon adenocarcinoma) in BALB/c mice could support suppressed tumor growth in treated animals compared with those injected with DOX alone (Bagheri et al., 2020). On the other hand, biomimetic nanovesicles derived from human-induced pluripotent stem cells (iPSCs)-derived MSCs loaded with DOX demonstrated more notable cytotoxic effects on doxorubicin-resistant triple-negative breast cancer (TNBC) cells, and also robustly reduced the occurrence and burden of metastases in murine models of TNBC in comparison with the free or liposomal doxorubicin application (Zhao et al., 2020).

MSC-Based Delivery of Prodrugs

Modified MSCs can express specific enzymes, in particular, HSV-thymidine kinase (TK) or cytosine deaminase (CD), converting inactive intravenously injected prodrugs, including ganciclovir (GCV) and 5-fluorocytosine (5-FC), into active cytotoxic agents, which in turn, can decrease systemic toxicity by easing tumor-localized chemotherapeutic activity (Pastorakova et al., 2020). Altogether, MSCs as an attractive carrier of therapeutic genes exemplify an exclusive device to stimulate drug activation within a neoplastic mass because of their property to home into tumors (Table 4).

TABLE 4

MSC-based delivery of pro-drugs for cancer therapy.

Cancer	Prodrug	Main results
Melanoma	5-FC	Marked inhibition of tumor growth with direct intratumoral transplantation of MSCs expressing CD followed by systemic injection of 5-FC in melanoma mice models (Krasikova et al., 2015)
Osteosarcoma	5-FC	Inhibition of the tumor growth in mice subcutaneously injected with osteosarcoma Cal72 cells following administration of stable CD/5-FC MSC cell line (NguyenThai et al., 2015)
Lewis lung carcinoma	5-FC	Significant inhibition of tumor growth with direct intratumoral transplantation of AT-MSCs expressing CD followed by systemic injection of 5-FC in mice models resulted in prolonged survival of treated mice (Krassikova et al., 2016)
Breast cancer	5-FC	Marked abrogation of tumor growth and attenuation of lung metastases with direct intratumoral transplantation of iPSC-derived MSCs expressing CD followed by systemic injection of 5-FC in breast tumor mice models (Ullah et al., 2017)
Glioma	5-FC	Potent inhibition of tumor development in glioma tumor-bearing mice following administration of MSC expressing CD with 5-FC (Chung et al., 2016)
Glioma	5-FC	The promoted lifespan and reduced tumor volume in C6 glioma-bearing rats by injection of MSC expressing CD with 5-FC (Fei et al., 2012)
Breast cancer	5-FC	Potent antitumor effects in SKOV3 tumor-bearing murine by administration of MSC expressing CD with 5-FC (Nouri et al., 2015)
Colorectal cancer	GCV	Inhibiting of tumor development and inducing tumor apoptosis in xenograft models established by injection of HT29 cells into the subcutaneous of right axilla of nude mice following injection of placental-MSCs expressing TK with GCV (Yang et al., 2019)
Glioma	GCV	Elimination of glioma cell lines and primary human glioblastoma cells following combination therapy with MSCs expressing TK and GCV in vitro (Pastorakova et al., 2020)
Cervical cancer	GCV	Reduction in tumor size along with improvement of the survival resulting from activation of NK cells and CTLs in tumor-bearing mice following administration of MSCs expressing TK with GCV (Kenarkoohi et al., 2020)
Glioma	GCV	Inhibition of tumor development in tumor-bearing mice following injection of MSCs expressing TK with GCV (Mori et al., 2010; Uchibori et al., 2009)
Hepatocellular carcinoma (HCC)	GCV	Marked inhibition of tumor growth in hepatic xenografts following administration of MSCs expressing TK with GCV (Niess et al., 2011)
Pancreatic cancer	GCV	Notable reduction in primary pancreatic tumor growth in xenograft models and reduced liver metastases following administration of MSCs expressing TK with GCV (Zischek et al., 2009)
Lung melanoma metastasis	GCV	Significant reduction in tumor colonization resulted in amelioration in the survival of murine melanoma lung metastasis models following injection of MSCs expressing TK with GCV (Zhang et al., 2015)

5-FC+CD

The 5-FC has been presented as a prodrug combined with the CD suicide gene. MSC-expressing CD (MSC-CD) have been formerly confirmed to suppress the development of subcutaneous prostate cancer xenografts upon converting the non-toxic 5-FC into the antineoplastic 5-fluorouracil (5-FU) (Abrate et al., 2014). Moreover, cytotoxicity of the MSC-CD on murine prostate tumor cells and also their marked tumor tropism has been documented. Importantly, systemic administration of the AT-MSC-CD followed by intraperitoneal injection of 5-FC could result in tumor deterioration in the transgenic adenocarcinoma of the mouse prostate (TRAMP) model (Abrate et al., 2014). In addition, it has been suggested that tumor cells stably transduced with CD gene by retrovirus infection could secret exosomes acting similarly like MSC-CD exosomes (Altanerova et al., 2021). These findings signify that cancer cell-derived suicide gene exosomes could home to their cells of origin, and thereby provide promising targeted therapy option for tumors (Altanerova et al., 2021). Furthermore, MSC-CD could stimulate remarkable anticancer effects with an extensive therapeutic index by converting 5-FC into 5-FU in glioblastoma multiforme (GBM) xenograft models (Chang et al., 2020). Besides, study of the potent antitumor effects of the MSC-CD in a combination regimen with temozolomide (TMZ), a well-known oral alkylating agent used to treat GBM, in vitro and a mouse orthotopic GBM model showed that TMZ and MSC-CD with 5-FC synergistically could hinder U87 glioma cell proliferation concurrently triggering cell cycle arrest and DNA breakage (Chang et al., 2020). On the other hand, administration of MSC-CD tracked by the serial treatment with 5-FC and TMZ showed more prominent suppressive effects on tumor development in animal models (Chang et al., 2020). Also, investigations in glioma C6 cells bearing murine models showed that transplantation of CD-expressing MSCs could attenuate tumor burden in proportion to 5-FC dosages (Chang et al., 2010). Nonetheless, single treatment was inadequate, and only succeeding transplantations showed the capability for delaying tumor growth, implying that total CD enzyme activity is a significant factor playing an influential role in determining the clinical efficacy for CD gene therapy (Chang et al., 2010). Another study in a model of nude mice bearing subcutaneous tumors of human gastric cancer MKN45 cells verified the therapeutic efficacy of BM-MSC-CD by intravenous route followed by systemic 5-FC treatment. Meanwhile, tumor mass and weights of mice infused with BM-MSC-CD were reduced considerably upon treatment with 5-FC, while mice treated with 5-FC alone did not experience any reduction in both tumor volume and bodyweights (You et al., 2009). Furthermore, assessment of the bystander effect presented that the MSC-CD increased human osteosarcoma Cal72 cytotoxicity in co-culture condition in the existence of 5-FC; on the other hand, MSC-CD injection along with 5-FC suppressed tumor development compared with control mice in Cal72 cell-bearing mice (NguyenThai et al., 2015).

Combination therapy with AT-MSCs expressing CD and lysomustine chemotherapy, a nitrosourea derivative of lysine, followed by systemic injection of 5-FC in mice bearing late-stage Lewis lung carcinoma (LLC) led to the significant attenuation of tumor growth, as evidence by decreased tumor volume (Krassikova et al., 2016). Moreover, the intervention resulted in prolonged survival of transplanted mice. More significantly, combination therapy showed a higher antitumor effect in murine models compared with monotherapies with injected AT-MSC-CD or lysomustine alone (Krassikova et al., 2016). Likewise, intratumoral transplantation of the iPS-derived MSCs expressing CD into a murine xenogeneic model of human breast cancer followed by 5-FC administration not only induced tumor regression but also diminished lung metastases (Ullah et al., 2017).

GCV+HSV-TK

Conceding findings, engineered MSC to express HSV/TK (MSC-HSV/TK) could phosphorylate GCV to its toxic metabolites and consequently induce tumor regression in human tumors (Kenarkoohi et al., 2020). In this regard, investigations have indicated that intratumoral administration of MSCs transduced with a lentivector expressing the HSV/TK followed by intraperitoneally administration of GCV could stimulate antitumor effects against a mouse cervical cancer model established by subcutaneous injection of TC-1 cells into female C57BL/6 mice (Kenarkoohi et al., 2020). The treatment caused a significant reduction in tumor mass, an improvement in survival, and also NK cell and cytotoxic T lymphocyte (CTL) antitumor activities, documenting the efficacy of MSC-expressing TK for cervical cancer therapy (Kenarkoohi et al., 2020). Similarly, HSV-TK-expressing UC-MSCs accompanied by GCV treatment exerted bystander cytotoxic effect on GBM U87 cells in vitro and also in U87-bearing murine models (Wei D. et al., 2019).

Wei D. et al. (2019) found that the ratio of MSC-TK and tumor cells could play an important role in determining the therapeutic effect rates, and a higher MSC-TK/U87 ratio led to a better effect. Besides, AT-MSC-TK elicited cytotoxic influences on human glioblastoma cells 8-MG-BA, 42-MG-BA, and U-118 MG followed by GCV treatment in vitro, in particular, on 8-MG-BA cells (Matuskova et al., 2010). In addition, the analysis revealed the establishing of gap junctions between AT-MSC and treated cell lines as a mechanism involved in bystander cytotoxicity, which supports MSC application for methods relying on the bystander effect (Matuskova et al., 2010). Besides, there are some evidence revealing that valproic acid (VPA) could promote the expression and activation of the gap junction proteins, such as connexin (Cx) 43 and 26, in glioma cells, and thereby ameliorate the bystander killing effects of the BM-MSC-TK and GCV treatment (Ryu et al., 2012). Combination therapy with VPA and BM-MSC-TK synergistically induced apoptosis in glioma cells by caspase activation in vitro and also potently suppressed tumor development and extended the survival of glioma-bearing mice (Ryu et al., 2012). Furthermore, studies in murine models of the metastatic renal cell carcinoma (RCC) demonstrated that MSCs engineered to co-express TRAIL and HSV-TK (MSC-TRAIL-TK) remained longer in the lungs of metastatic xenograft models in comparison with the parental MSCs (Kim et al., 2013). Notably, MSC-TRAIL-TK injection followed by GCV treatment declined tumor nodule frequencies in the lungs more obviously than MSC-TRAIL or MSC-TK, which in turn resulted in prolonged survival. Furthermore, it was found that the antimetastatic influences of MSC-TRAIL-TK were enriched through serial injections but not via augmented dose, offering an attractive therapeutic tactic to eliminate metastatic tumors by repeated injection of MSC-TRAIL-TK (Kim et al., 2013). In consistent with other findings, engineered human placenta-derived MSCs (hP-MSCs) to express HSV-TK showed antitumor effects against colon cancer xenograft models established by subcutaneous administration of HT29 cells into nude mice. According to the results, the injection of the engineered hP-MSCs considerably suppressed tumor growth, while the observed effect was improved by GCV treatment (Yang et al., 2019). Moreover, in vivo reports have described that MSC-TK could effectively home into primary pancreatic tumor stroma and subsequently induce CCL5 promoter activation, as evidenced by the use of enhanced green fluorescent protein (eGFP)- and red fluorescent protein (RFP)-reporter genes, in C57/Bl6 models (Zischek et al., 2009). Furthermore, upon treatment with GCV, modified MSCs triggered a significant inhibitory effect on primary pancreatic tumor growth as well as on occurrences of metastases (Zischek et al., 2009). Furthermore, the safety, tolerability, and efficacy of treatment with MSC-TK combined with GCV have been evidenced by the accomplishment of phase 1 clinical trial on s participants suffering from advanced gastrointestinal cancer carried out by von Einem and his coworkers between November 2013 and December 2014. During follow-up, the intervention led to stable disease in 4/6 patients, and progressive disease in 2/6 patients in the absence of any serious treatment-related events, verifying the acceptable safety and tolerability in enrolled patients with advanced gastrointestinal cancer (von Einem et al., 2017).

MSC-Based Delivery of Trail

In spite of potential pathological conditions, apoptosis procedure can be a capable target for tumor therapy. TRAIL, or Apo-2 ligand (Apo2L), is a member of TNF cytokine superfamily with attractive potent anticancer function owing to its unique competence to target cancerous cells without any harm to adjacent normal cells. This is because the expression of TRAIL-specific receptors, termed as death receptors (DR), are significantly higher in cancer cells compared with normal cells (Figure 3; Nagane et al., 2001; Lemke et al., 2014; Yamamoto et al., 2020).

FIGURE 3

The mechanism of TRAIL-induced apoptosis in tumor cells. TRAIL typically elicits apoptosis in target cells following binding to its specific receptors, known as DRs, leading to the activation of the caspase cascade. TRAIL/Apo2L, tumor necrosis factor (TNF)-related apoptosis-inducing ligand; DR, death receptor; BAX, BCL2-associated X; BAK1, BCL2 antagonist/killer; FADD, Fas-associated via death domain; tBid, truncated BH3 interacting domain death agonist.

In the context of cancer’s cell-based therapy, TRAIL can be utilized either in its natural shape as a full-length and membrane-bound protein (FL-TRAIL) or as a modified shape generally termed as soluble TRAIL (sTRAIL) (Mielczarek-Palacz et al., 2020; Patrick et al., 2020) (Table 5). 89Zr-oxine labeling and positron emission tomography (PET)/computed tomography (CT) imaging for tracking MSC-TRAIL biodistribution in metastatic lung adenocarcinoma xenograft models evidenced the effective delivery of MSC-TRAIL to the lungs in a murine model up to 1-week postinjection, verified through in vivo bioluminescence imaging, autoradiography, and fluorescence imaging on tissue sections (Patrick et al., 2020). MSC-sTRAIL could dramatically stimulate more powerful apoptosis-inducing activity compared with cells showing FL-TRAIL. TRAIL although could motivate the expression of prometastatic molecules, such as CXCL5/ENA-78 and IL-6 in prostate cancer cells, this influence could be defeated by combination therapy with an AKT inhibitor (Mohr et al., 2019). Accordingly, combination therapy using small-molecule drugs and MSC-sTRAIL could sensitized tumor cells to TRAIL, and also decreased the concerns of side-effect-initiating cytokine production (Mohr et al., 2019). Conversely, another report has signified that MSC-FL-TRAIL could induce more powerful cytotoxicity against cancer cells than MSC-sTRAIL, and also could defeat cancer cell resistance to recombinant TRAIL, thereby suggesting that MSC delivery of FL-TAIL is superior to MSC delivery of sTRAIL for cancer therapy (Yuan et al., 2015). Besides, AT-MSCs modified to express sTRAIL were found to induce apoptosis in pancreatic ductal adenocarcinoma (PDAC) BxPC-3 and MIA PaCa-2 cell lines, and also in primary PDAC cells (Spano et al., 2019). On the other hand, sTRAIL secreted by AT-MSCs relocated into the tumor stroma and then remarkably negatively modified tumor development in vivo with a substantial decrease in tumor size and exerting antiangiogenic effect (Spano et al., 2019).

TABLE 5

MSC-based delivery of TRAIL for cancer therapy.

Cancer	Main results
Neuroblastoma	Efficient migration to tumor tissue resulted in delayed tumor proliferation in xenografts (Nieddu et al., 2019)
Lung carcinoma	Significant abrogation of tumor growth in A549 cell-bearing mice (Mohr et al., 2008; Yan et al., 2016)
Breast cancer	Inducing the apoptosis of MCF-7 cells in vitro (Khorashadizadeh et al., 2015)
Several tumors	Abrogation of breast, lung, and cervical cancer cell growth in vitro and in vivo (Loebinger et al., 2009)
Melanoma	Suppressing tumor growth in B16F0 cells bearing murine (Salmasi et al., 2020)
Hepatocellular carcinoma	Stimulating apoptosis in N1-S1, HepG2, and MHCC97-H cell line in vitro and in vivo by upregulation of caspase-3, and death receptors 4 (DR4) and 5 (DR5) expression (Deng et al., 2014)
Glioma	Delayed tumor growth resulted in prolonged survival in U87MG cell-bearing mice (Yang et al., 2009; Xia et al., 2015)
Mesothelioma	Delayed tumor growth in HMESO cell xenografts (Lathrop et al., 2015)
Sarcoma	Inhibition of tumor cell proliferation in tumor-bearing mice by inducing the caspase activation (Guiho et al., 2016)
Colorectal cancer	Stimulation of the antitumor effects by MSCs-TRAIL plus 5-FU on HCT116 cell-bearing nude mice (Yu et al., 2013)
Brain metastatic breast cancer	Tumor cell apoptosis by MSCs genetically modified to co-overexpress CXCR4 and TRAIL in MDA-MB-23-bearing mice (Liu M. et al., 2020)
Esophageal cancer	Inhibition of tumor growth in vitro and in Eca-109 cell-bearing mice (Li et al., 2014)
Pancreatic cancer Glioblastoma	Induced antitumor effects of paclitaxel primed MSC-TRAIL on CFPAC-1 and U87-MG cell line in vitro (Coccè et al., 2020)
Multiple myeloma	Specific induction of myeloma U-266 cells apoptosis in vitro and in vivo by caspase-3 activation leading to reduced tumor burden in xenografts (Cafforio et al., 2017)

Tumor necrosis factor-related apoptosis-inducing ligand delivery by MSC-derived extracellular vesicles (EVs) is considered an effective antitumor treatment option. In this regard, EVs derived from MSC-TRAIL showed cytotoxicity against lung cancer lines (A549, NCI-H460, and NCI-H727), malignant pleural mesothelioma lines (H2795, H2804, H2810, and H2818), renal cancer lines (RCC10 and HA7-RCC), human breast adenocarcinoma line (MDAMB231; M231), and neuroblastoma line (SHEP-TET) without any cytotoxicity against primary human bronchial epithelial cells (Yuan et al., 2017). Importantly, EVs derived from MSC-sTRAIL stimulated evident apoptosis in TRAIL-resistant cancer cells which were improved using a CDK9 inhibitor, thus suggesting MSC-derived EVs as an eligible anticancer therapy (Yuan et al., 2017).

Similarly, injection of the MSC-sTRAIL intravenously or intraperitoneally into the xenograft model of mesothelioma supported tumor regression concurrently decreased local inflammation. These proof-of-concept educations propose that MSC-sTRAIL could be beneficial toward malignant mesothelioma (Lathrop et al., 2015). Besides, targeting of CD133-positive cancer stem cells (CSCs) by MSC-TRAIL showed a prospective role of modifying apoptosis-related genes in non-small cell lung cancer (NSCLC) (Fakiruddin et al., 2019). Regardless of maintaining their multipotent property, MSC-TRAIL exposed with CSCs led to the significant mitigation in their proliferation and also induced tumor cell apoptosis in vitro largely by activation of the apoptosis intrinsic pathway in CSCs (Fakiruddin et al., 2019). Molecular analysis exemplified that modifying the expression of NF-κB1, BAG cochaperone 3 (BAG3), MCL1, growth arrest, DNA damage-inducible alpha (GADD45A), and harakiri (HRK) was responsible for observed anticancer effects exerted by MSC-TRAIL in CSCs (Fakiruddin et al., 2019).

Conclusion and Prospect

Mesenchymal stem/stromal cells are a capable and attractive therapeutic option for cancer therapy, ranging from solid tumors to hematological malignancies. The engineered MSCs have unique competence to specifically affect malignant cells without notable untoward effects and systemic toxicity. Nonetheless, as cancerous cells and MSCs interrelate in different ways, either suppressing or sustaining tumor development concerning multiple factors may arise from this interaction. Based on the literature, MSCs could sustain tumor development by a myriad of mechanisms, encompassing promotion of drug resistance, pro-angiogenic activities, and induction of metastasis process by triggering epithelial-mesenchymal transition (EMT) concomitant with enrichments of the CSC niche (Figure 4). MSC-secreted factors are responsible for the cited effects which affect multiple hallmarks of cancer. However, homing aptitude of MSCs makes them dependable nominees as drug delivery vehicles for therapeutic agents. Further studies respecting the tumor growth mechanisms of MSCs may increase the possibility to utilize them for cancer treatment by amelioration of their preparation procedure to attenuate the risk of tumor cell growth. In addition, a more comprehensive knowledge of the particular molecular mechanisms contributing to the protumorigenic functions of MSCs is of paramount importance. In this regard, an alternative strategy for utilizing the intact MSCs for antitumor agent delivery is the employment of an MSC-derived conditioned medium, minimizing the risk of tumorigenicity in MSC-based approach for cancer treatment. In sum, it seems that weakening the MSC tumor-supportive activities by various strategies such as combination therapy with small molecules (e.g., tyrosine kinase inhibitors) may enable achievement of the more desired therapeutic outcomes.

FIGURE 4

MSC-induced tumor-supportive processes. MSCs could stimulate tumor-supportive procedures, such as chemoresistance, metastasis, and angiogenesis by a myriad of mechanisms, such as differentiation into CAFs, CSCs, and pericytes, promotion of EPC recruitment into the tumor area, inhibition of the chemotherapeutic drug’s functions, and also triggering of EMT process. MSCs, mesenchymal stem/stromal cells; CSCs, cancer stem cells; EPCs, endothelial progenitor cells; CAFs, cancer-associated fibroblasts; EMT, epithelial-mesenchymal transition.

Author Contributions

All authors contributed to the conception and the main idea of the work and read and approved the final version of the work to be published. AH, AA, HR, MS, DB, WA, FM, MZ, YY, and MY drafted the main text, figures, and tables. MJ supervised the work and provided the comments and additional scientific information. MC and YP also reviewed and revised the text.

Conflict of Interest

The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.